The goal of this proposal is to continue the product development of a novel vaccine approach that incorporates nucleoprotein (NP) as a conserved internal component of influenza virus linked to an immunostimulatory oligonucleotide to induce protective immunity against divergent and potentially pandemic influenza strains. Specifically, the plan is to complete process development and scale up for GMP production of recombinant NP in an E. coli system. Process development, scale-up and GMP production of the NP-ISS conjugate for the vaccine will then be performed, as well as IND enabling toxicology and stability studies. In addition, characterization of the immunogenicity and protective immunity provided by the NP-ISS vaccine alone and NP-ISS combined with standard trivalent inactivated vaccine will be performed in animal models. The Specific Aims of the proposal are: [unreadable] 1. Produce GMP-grade NP-ISS conjugate [unreadable] * GMP manufacture of recombinant nucleoprotein in the E. coli production system [unreadable] * Process development and scale-up of NP-ISS conjugate formulation [unreadable] * GMP manufacture of NP-ISS formulation [unreadable] 2. Perform IND enabling toxicology on NP-ISS vaccine [unreadable] TIV [unreadable] 3. Perform further characterization of the NP-ISS vaccine. [unreadable] * Assess protection provided by E. coli derived NP-ISS in the mouse challenge model against drift (H1), shift (H3) and avian (H5) influenza strains. [unreadable] * Assess protection following immunization in the ferret influenza challenge model with drift (H1),shift (H3) and avian (H5) influenza strains. [unreadable] * Optimize the combination of NP-ISS and TIV in primates to allow HA dose sparing while inducing NP-specific cell mediated immunity. [unreadable] * Evaluate immunogenicity of the NP-ISS vaccine combined with trivalent inactivated vaccine in elderly mice. [unreadable] [unreadable] [unreadable] [unreadable]